Principles of microscopy

1. Magnification – number of times an object is enlarged

2. Contrast – difference between background and the specimen
Ex: wet mount of colorless cells (like bacteria) in a dark background
3. Resolution – also known as resolving power
- is the capacity to distinguish 2 points as separate
Ex: 0.2 µm resolution means cells as far as 0.2 µm can be seen as 2 separate cells, but if
- distance is below 0.2 µm between cells, the cells will merge as one cell.
- Also cells smaller as 0.2 µm, will be seen as one cell also

Diffraction- capacity of bending of light

Refraction – capacity of light particles to scatter and separate
We use cedar wood oil or cedar tree oil, placed on top of the cover slip for OIO (oil immersion objective)
to prevent scattering of light particles as such particles pass thru the slide and specimen and exits thru
the eye piece( both diffraction & refraction refers to bending of light. But refraction looks at light
particles to scatter)

objective x eyepiece magnification = Total magnification. The given magnification values are from the
Olympus compound light microscope we are using.

Objectives Magnification Eyepiece Total Magnification

scanner 4X 10 X 40X
LPO 10X 10X 100X
HPO 40X 10X 400X
OIO 100X 10X 1000X

Eyepiece: has magnification of 10X

10X means an object is magnified 10 times
So 1000X in OIO total magnification values means, an object is magnified 1000 times
Light source: mirror or electric type ( push the on button first)

Reminders
-start by using the lowest objective upon viewing (scanner) before jumping to other objectives with
higher magnification values
-You will not be able to view the details in OIO if the image was not properly refined using the lower
objectives, resulting in a blurry image, go back to objectives with lower values of magnification ( LPO)
-upon using microscope,check 1.power source then 2.light source then 3.regulate light intensity by iris
diaphragm
-carry microscope with 2 hands (1 is at the arm, 1 is at the base)
-view ocular lenses with 2 eyes
-clean microscope before and after using (ocular+objective lenses+stage) with: lens cleaner or alcohol
-upon returning, rotate the nosepiece to the lowest objective (scanner) in focus

How not to break glass slides:- scanner & LPO can be used with coarse adjustment, HPO and OIO can
only be used with fine adjustment. HPO and OIO objectives will almost touch the stage. Only fine
adjustment is used to refine the details of cells upon viewing the microscope.